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An optimised protocol for detection of SARS-CoV-2 in stool
Tianqi Li1,2, Enriqueta Garcia-Gutierrez1, Daniel A Yara1
1Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
BMC Microbiology
|September 7, 2021
Summary
A new sensitive method reliably detects SARS-CoV-2 in stool samples, crucial for understanding gut persistence and ensuring the safety of fecal microbiota transplants.
Area of Science:
- Virology
- Gastroenterology
- Molecular Diagnostics
Background:
- SARS-CoV-2 detected in stool suggests fecal-oral transmission potential.
- Detecting SARS-CoV-2 in stool is challenging due to the complex matrix.
- Reliable methods are needed for gut viral persistence studies and fecal microbiota transplant (FMT) safety.
Purpose of the Study:
- To develop a sensitive and reliable method for SARS-CoV-2 detection in stool samples.
- To address the unmet need for accurate viral detection in complex fecal matrices.
Main Methods:
- Inactivated SARS-CoV-2 spiked into stool samples at known concentrations.
- Viral particle concentration via ultrafiltration.
- RNA extraction followed by real-time reverse-transcription polymerase chain reaction (RT-qPCR) using CDC primers.
Main Results:
- The developed method detected SARS-CoV-2 in most tested stool samples.
- A limit of detection as low as 50 viral particles per 100 mg of stool was achieved.
- The N1 primer set demonstrated superior reliability and precision, achieving a limit of detection of 1 viral particle per mg of stool.
Conclusions:
- A sensitive method for SARS-CoV-2 stool detection is established.
- This method supports the assessment of viral persistence in the gut.
- The method enhances the safety of clinical practices like FMT.
